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Objective: To evaluate the effect of point of care ultrasonography (POCUS) performed for heart, lung, aorta, hepatobiliary and deep veins on the diagnosis, length of stay (LOS) in emergency department (ED) and cost in patients admitted to the ED with chest pain., Study Design: Prospective randomised controlled, parallel-group trial., Place and Duration of Study: Sakarya University Training and Research Hospital, Sakarya Turkey, from September 2018 to March 2019., Methodology: Patients (≥18 years) with chest pain were randomly assigned at a 1:1 ratio to a standard diagnostic strategy (control group) or to standard diagnostic strategy supplemented with POCUS (POCUS group). Data obtained from the study were analysed using IBM SPSS Statistics 21., Results: Two hundred and eight patients were randomly assigned to the control (n=104) and POCUS groups (n=104), respectively. The mean age was 50.42 ± 16.15, and 54% were men. The most common comorbidity was hypertension (43%). Non-ST elevation myocardial infarction and musculoskeletal pain were the most common presumptive diagnoses. POCUS significantly reduced the LOS in ED. Detection of pathology in the POCUS increased the rate of hospitalisation. In addition, POCUS significantly shortened the LOS in the ED in patients who were discharged. The median LOS in the ED for the POCUS group was 133 min (91-279), which was significantly shorter than that of the control group at 215 min (118-372) (p=0.006). Although the average costs were also reduced, the difference was not statistically significant (p=0.269)., Conclusion: POCUS is a repeatable, practical imaging method which does not require radiation, reduces LOS in the ED statistically significant. However, further studies are needed to determine its usefulness in the ED. Key Words: Chest pain, Cost, Emergency medicine, Length of stay, Point of care ultrasound.

Tantalum possesses remarkable chemical and mechanical properties, and thus it is considered to be one of the next generation implant materials. However, the biological properties of tantalum remain to be improved for its use in tissue engineering applications. To enhance its cellular interactions, implants made of tantalum could be modified to obtain nanofeatured surfaces via the electrochemical anodization process. In this study, anodization parameters were adjusted to obtain a nanoporous surface morphology on tantalum surfaces and systematically altered to control the pore sizes from 25 to 65 nm using an aqueous HF:H 2 SO 4 electrolyte. Results indicated the formation of Ta 2 O 5 -based nanoporous surface layers, which had up to 28% more surface area and increased nanophase roughness (more than twofolds) compared to nonporous tantalum upon the anodization. It was observed that the nanoporous tantalum oxide surfaces promoted nearly 25% more fibroblast proliferation at 5 days in vitro and 15.5% more cellular spreading. Thus, nanoporous tantalum oxide surfaces can be used to increase biological interactions of the cells and provide a means of improving bioactivity of tantalum for biomaterial applications., (© 2020 Wiley Periodicals, Inc.)

Exosomes derived from mesenchymal stem cells are extracellular vesicles released to facilitate cell communication and function. Recently, polylactic acid (PLA), calcium silicates (CaSi), and dicalcium phosphate dihydrate (DCPD) have been used to produce bioresorbable functional mineral-doped porous scaffolds-through thermally induced phase separation technique, as materials for bone regeneration. The aim of this study was to investigate the effect of mineral-doped PLA-based porous scaffolds enriched with exosome vesicles (EVs) on osteogenic commitment of human adipose mesenchymal stem cells (hAD-MSCs). Two different mineral-doped scaffolds were produced: PLA-10CaSi-10DCPD and PLA-5CaSi-5DCPD. Scaffolds surface micromorphology was investigated by ESEM-EDX before and after 28 days immersion in simulated body fluid (HBSS). Exosomes were deposited on the surface of the scaffolds and the effect of exosome-enriched scaffolds on osteogenic commitment of hAD-MSCs cultured in proximity of the scaffolds has been evaluated by real time PCR. In addition, the biocompatibility was evaluated by direct-contact seeding hAD-MSCs on scaffolds surface-using MTT viability test. In both formulations, ESEM showed pores similar in shape (circular and elliptic) and size (from 10-30 µm diameter). The porosity of the scaffolds decreased after 28 days immersion in simulated body fluid. Mineral-doped scaffolds showed a dynamic surface and created a suitable bone-forming microenvironment. The presence of the mineral fillers increased the osteogenic commitment of hAD-MSCs. Exosomes were easily entrapped on the surface of the scaffolds and their presence improved gene expression of major markers of osteogenesis such as collagen type I, osteopontin, osteonectin, osteocalcin. The experimental scaffolds enriched with exosomes, in particular PLA-10CaSi-10DCPD, increased the osteogenic commitment of MSCs. In conclusion, the enrichment of bioresorbable functional scaffolds with exosomes is confirmed as a potential strategy to improve bone regeneration procedures., Competing Interests: The authors declare no conflict of interest.

An increasing interest in the fabrication of implants made of titanium and its alloys results from their capacity to be integrated into the bone system. This integration is facilitated by different modifications of the implant surface. Here, we assessed the bioactivity of amorphous titania nanoporous and nanotubular coatings (TNTs), produced by electrochemical oxidation of Ti6Al4V orthopedic implants' surface. The chemical composition and microstructure of TNT layers was analyzed by X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). To increase their antimicrobial activity, TNT coatings were enriched with silver nanoparticles (AgNPs) with the chemical vapor deposition (CVD) method and tested against various bacterial and fungal strains for their ability to form a biofilm. The biointegrity and anti-inflammatory properties of these layers were assessed with the use of fibroblast, osteoblast, and macrophage cell lines. To assess and exclude potential genotoxicity issues of the fabricated systems, a mutation reversal test was performed (Ames Assay MPF, OECD TG 471), showing that none of the TNT coatings released mutagenic substances in long-term incubation experiments. The thorough analysis performed in this study indicates that the TNT5 and TNT5/AgNPs coatings (TNT5-the layer obtained upon applying a 5 V potential) present the most suitable physicochemical and biological properties for their potential use in the fabrication of implants for orthopedics. For this reason, their mechanical properties were measured to obtain full system characteristics., Competing Interests: The authors declare no conflict of interest.

Cholesterol is a major structural component of the plasma membrane (PM). The majority of PM cholesterol forms complexes with other PM lipids, making it inaccessible for intracellular transport. Transition of PM cholesterol between accessible and inaccessible pools maintains cellular homeostasis, but how cells monitor the accessibility of PM cholesterol remains unclear. We show that endoplasmic reticulum (ER)-anchored lipid transfer proteins, the GRAMD1s, sense and transport accessible PM cholesterol to the ER. GRAMD1s bind to one another and populate ER-PM contacts by sensing a transient expansion of the accessible pool of PM cholesterol via their GRAM domains. They then facilitate the transport of this cholesterol via their StART-like domains. Cells that lack all three GRAMD1s exhibit striking expansion of the accessible pool of PM cholesterol as a result of less efficient PM to ER transport of accessible cholesterol. Thus, GRAMD1s facilitate the movement of accessible PM cholesterol to the ER in order to counteract an acute increase of PM cholesterol, thereby activating non-vesicular cholesterol transport., Competing Interests: TN, BE, LK, AT, DK, FW, KT, FT, MW, YS No competing interests declared, (© 2019, Naito et al.)

The outer membrane (OM) of Gram-negative bacteria is a permeability barrier that impedes the entry of external insults, such as antibiotics and bile salts. This barrier function depends critically on the asymmetric lipid distribution across the bilayer, with lipopolysaccharides (LPS) facing outside and phospholipids (PLs) facing inside. In Escherichia coli, the OmpC-Mla system is believed to maintain OM lipid asymmetry by removing surface exposed PLs and shuttling them back to the inner membrane (IM). How proteins in the pathway interact to mediate PL transport across the periplasm is not known. Evidence for direct transfer of PLs between these proteins is also lacking. In this study, we mapped the interaction surfaces between the two PL-binding proteins, MlaC and MlaD, using site-specific in vivo photo-cross-linking, and obtained a physical picture for how these proteins may transfer PLs. Furthermore, we demonstrated using purified proteins that MlaD spontaneously transfers PLs to MlaC, suggesting that the latter has a higher affinity for PLs. Our work provides insights into the mechanism of bacterial intermembrane lipid transport important for the maintenance of OM lipid asymmetry.

Objective: Prostate specific antigen (PSA) with digital rectal examination is used for diagnosis of prostate cancer (PCa), where definite diagnosis can only be made by prostate biopsy. Recently neutrophil gelatinase-associated lipocalin (NGAL), a lipocalin family member glycoprotein, come into prominence as a cancer biomarker. This study is aimed to test serum NGAL as a diagnostic biomarker for PCa and discriminate PCa from benign prostatic hyperplasia (BPH)., Material and Methods: In this prospective study, 90 patients who underwent transrectal ultrasound-guided 12-core prostate biopsy between May 2015 and September 2015, were evaluated. Histopathologically diagnosed 45 PCa and 45 BPH patients were enrolled in this study. Serum NGAL and PSA levels of all participants were measured, then these data were evaluated by statistical programs., Results: When sensitivity fixed to 80% specificity of NGAL was better than PSA (49%, 31% respectively). Receiver operating characteristic (ROC) curve analysis showed that NGAL alone or its combined use with PSA have better area under curve (AUC) results than PSA alone (0.662, 0.693, and 0.623 respectively)., Conclusion: In conclusion NGAL gave promising results such as increased sensitivity and a better AUC values in order to distinguish PCa from BPH. NGAL showed a potential to be a non-invasive biomarker which may decrease the number of unnecessary biopsies., Competing Interests: Conflict of Interest: No conflict of interest was declared by the authors.

In Gram-negative bacteria, lipid asymmetry is critical for the function of the outer membrane (OM) as a selective permeability barrier, but how it is established and maintained is poorly understood. Here, we characterize a non-canonical ATP-binding cassette (ABC) transporter in Escherichia coli that provides energy for maintaining OM lipid asymmetry via the transport of aberrantly localized phospholipids (PLs) from the OM to the inner membrane (IM). We establish that the transporter comprises canonical components, MlaF and MlaE, and auxiliary proteins, MlaD and MlaB, of previously unknown functions. We further demonstrate that MlaD forms extremely stable hexamers within the complex, functions in substrate binding with strong affinity for PLs, and modulates ATP hydrolytic activity. In addition, MlaB plays critical roles in both the assembly and activity of the transporter. Our work provides mechanistic insights into how the MlaFEDB complex participates in ensuring active retrograde PL transport to maintain OM lipid asymmetry., Competing Interests: The authors declare that no competing interests exist.

Silk possesses many beneficial wound healing properties, and electrospun scaffolds are especially applicable for skin applications, due to their smaller interstices and higher surface areas. However, purified silk promotes microbial growth. Selenium nanoparticles have shown excellent antibacterial properties and are a novel antimicrobial chemistry. Here, electrospun silk scaffolds were doped with selenium nanoparticles to impart antibacterial properties to the silk scaffolds. Results showed significantly improved bacterial inhibition and mild improvement in human dermal fibroblast metabolic activity. These results suggest that the addition of selenium nanoparticles to electrospun silk is a promising approach to improve wound healing with reduced infection, without relying on antibiotics.

Catheter-associated infections, most of which are caused by microbial biofilms, are still a serious issue in healthcare and are associated with significant morbidity, mortality, and excessive medical costs. Currently, the use of nanostructured materials, especially materials with nanofeatured topographies, which have more surface area, altered surface energy, enhanced select protein adsorption, and selectively increased desirable cell functions while simultaneously decreasing competitive cell functions, seem to be among the most promising ways for reducing initial bacteria attachment, biofilm formation, and infections. In this study, polydimethylsiloxane (PDMS), a commonly used polymeric catheter material, was formulated to mimic the nanopatterned topography of natural tissue by using a template method with nanotubular anodized titanium. Results showed that increased PDMS surface nanoscale roughness alone can inhibit both Gram-negative ( Escherichia coli ) and Gram-positive ( Staphylococcus aureus ) bacteria adhesion and growth for up to 2 days, the time length of the current study. Additionally, increased fibroblast and endothelial cell adhesion on nano-PDMS indicated that this nanoscale topography had no toxic effects toward mammalian cells. Mechanistically, this study also developed a model for the first time to correlate bacteria responses to nanoscale roughness with initial protein and biomolecule adsorption (specifically, casein protein and glucose, which are unique biomolecules that mediate bacteria functions). Data revealed that the increase in nanoscale roughness and associated energy contributed to greater select casein adsorption during the first several minutes of culture, which is critical for decreasing bacteria attachment and growth. In contrast, no significant differences for glucose adsorption between samples before and after nanofabrication were identified. These results together indicated that the present biomimetic nanopatterned PDMS surface without any chemical or antibiotic modification has the potential to combat catheter-associated infections and should be further investigated.

The principle of guided bone regeneration (GBR) in orthopedic, cranio-maxillofacial and dental tissue engineering applications is to create a secluded space for the treatment of large bone defects while excluding fibrous connective tissue formation at the defect area. In dental surgeries, a GBR membrane is placed near the dental implant in post-extraction sockets to grow new bone at the implant site, along with inhibiting infection due to the microbial nature of the mouth flora. Poly[(R)-3-hydroxybutyric acid] (PHB) is a natural polyester synthesized by a wide variety of microorganisms which has been proposed for various biomedical applications. In this study, to improve the performance of PHB as a GBR, a NaOH based alkaline treatment was designed to create nanofeatured PHB membranes. The newly fabricated nanofeatured PHB membranes were investigated for GBR applications. The results showed that a quick, simple, and inexpensive sodium hydroxide treatment modified the nanostructured surface morphology and chemistry of the PHB membranes by inducing hydrolysis of the ester bonds in the PHB backbone creating carboxylic surface functional groups, which increased the hydrophilicity of the PHB surfaces. Cytocompatibility studies showed increased proliferation of human osteoblasts (bone forming cells) on the NaOH treated PHB membranes compared to the untreated ones. Importantly, in vitro bacterial studies with Staphylococcus aureus (S. aureus) indicated that the NaOH-treated PHB surfaces inhibited S. aureus growth more than 60% after 48 hours of culture compared to the untreated PHB membrane. Thus, this study, for the first time, showed that nanofeatured PHB membranes modified with a NaOH treatment may be a useful anti-bacterial, osteoconductive GBR membrane for numerous orthopedic, cranio-maxillofacial and dental tissue engineering applications.

Magnesium plays an important role in the body, mediating cell-extracellular matrix interactions and bone apatite structure and density. This study investigated, for the first time, the effects of adding magnesium oxide (MgO) nanoparticles to poly (l-lactic acid) (PLLA) and to hydroxyapatite (HA) nanoparticle-PLLA composites for orthopedic tissue engineering applications. Results showed that MgO nanoparticles significantly enhanced osteoblast adhesion and proliferation on HA-PLLA nanocomposites while maintaining mechanical properties (Young's modulus ∼1,000 MPa) suitable for cancellous bone applications. Additionally, osteoblasts (or bone-forming cells) cultured in the supernatant of degrading nanocomposites showed improved proliferation in the presence of magnesium, indicating that the increased alkalinity of solutions containing MgO nanocomposites had no toxic effects towards cells. These results together indicated the promise of further studying MgO nanoparticles as additive materials to polymers to enhance the integration of implanted biomaterials with bone., (Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

As an effort to create the next generation of improved skin graft materials, in this study, we modified the surfaces of a previously investigated material, silk fibroin, using a NaOH alkaline treatment to obtain a biologically inspired nanofeatured surface morphology. Such surfaces were characterized for roughness, energy, and chemistry. In addition, keratinocyte (skin-forming cells) adhesion and proliferation on such nanofeatured silk fibroin wound dressings were studied in an initial attempt to determine the promotion of an epidermal cover on the wound bed to form a new epidermal barrier. Dermal fibroblast adhesion and proliferation were also studied to assess the ability of nanostructured silk fibroin to replace damaged dermal tissue in chronic wounds (i.e., for diabetic foot ulcers). Results demonstrated for the first time that keratinocyte and fibroblast cell density was greater on nanofeatured silk fibroin membranes compared with non-treated silk fibroin surfaces. The enhancement in cellular functions was correlated with an increase in silk surface nanotopography, wettability and change in chemistry after NaOH treatment. Due to the present promising results, the newly developed nanofeatured silk fibroin membranes are exciting alternative skin graft materials which should be further studied for various skin patch and wound dressing applications., (© 2014 Wiley Periodicals, Inc.)

Major issues faced with the use of today's skin grafts are infection, scar tissue formation, insufficient keratinocyte (or skin producing cells) proliferation and high production costs. To overcome these limitations, we propose here for the first time, a nanofeatured poly(lactide-co-glycolide) (PLGA) membrane as a next generation antibacterial skin graft material. An alkaline surface treatment method was used to create random nanofeatures on PLGA membranes where sodium hydroxide (NaOH) concentration and exposure times were altered to control surface morphology. Most significantly, and without the use of antibiotics, results showed a decrease in Staphylococcus aureus (a dangerous pathogen infecting skin grafts) growth for up to ∼40% after 2 days of culture on nanofeatured PLGA membranes compared to untreated controls. Results also showed that while bacteria growth was stunted, mammalian cell growth was not. Specifically, cell culture results showed an increase in human epidermal keratinocyte density, while the density of scar tissue forming human dermal fibroblasts, did not change on nanofeatured PLGA surfaces compared to the untreated controls after 3 days of culture. These findings indicate that the alkaline treatment of PLGA membranes is a promising quick and effective manner to limit scar tissue formation and bacterial invasion while increasing skin cell proliferation for improving numerous wound-healing applications., (© 2014 Wiley Periodicals, Inc.)

In this study, 316L stainless steel with tunable nanometer pit sizes (0, 25, 50, and 60 nm) were fabricated by an anodization procedure in an ethylene glycol electrolyte solution containing 5 vol % perchloric acid. The surface morphology and elemental composition of the 316L stainless steel were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS). The nano-pit arrays on all of the 316L stainless steel samples were in a regular arrangement. The surface properties of the 316L stainless steel nano-pit surface showed improved wettability properties as compared with the untreated 316L stainless steel, as demonstrated by the lower contact angles which dropped from 83.0° to 28.6 to 45.4°. The anodized 316L stainless steel surfaces with 50 nm and 60 nm diameter pits were also more rough at the nanoscale. According to MTT assays, compared with unanodized (that is, nano-smooth) surfaces, the 50 and 60 nm diameter nano-pit surfaces dramatically enhanced initial human dermal fibroblast attachment and growth for up to 3 days in culture. Mechanistically, this study also provided the first evidence of greater select protein adsorption (specifically, vitronectin and fibronectin which have been shown to enhance fibroblast adhesion) on the anodized 316L stainless steel compared with unanodized stainless steel. Such nano-pit surfaces can be designed to support fibroblast growth and, thus, improve the use of 316L stainless steel for various implant applications (such as for enhanced skin healing for amputee devices and for percutaneous implants)., (© 2014 Wiley Periodicals, Inc.)

We demonstrate a 3x1 fiber-based photonic lantern spatial-multiplexer with mode-selectivity greater than 6 dB and transmission loss of less than 0.3 dB. The total insertion loss of the mode-selective multiplexers when coupled to a graded-index few-mode fiber was < 2 dB. These mode multiplexers showed mode-dependent loss below 0.5 dB. To our knowledge these are the lowest insertion and mode-dependent loss devices, which are also fully compatible with conventional few-mode fiber technology and broadband operation.

Infection of titanium (Ti)-based orthopedic implants is a growing problem due to the ability of bacteria to develop a resistance to today's antibiotics. As an attempt to develop a new strategy to combat bacteria functions, Ti was anodized in the present study to possess different diameters of nanotubes. It is reported here for the first time that Ti anodized to possess 20 nm tubes then followed by heat treatment to remove fluorine deposited from the HF anodization electrolyte solution significantly reduced both S. aureus and S. epidermidis growth compared to unanodized Ti controls. It was further found that the sterilization method used for both anodized nanotubular Ti and conventional Ti played an important role in the degree of bacteria growth on these substrates. Overall, UV light and ethanol sterilized samples decreased bacteria growth, while autoclaving resulted in the highest amount of bacteria growth. In summary, this study indicated that through a simple and inexpensive process, Ti can be anodized to possess 20 nm tubes that no matter how sterilized (UV light, ethanol soaking, or autoclaving) reduces bacteria growth and, thus, shows great promise as an antibacterial implant material., (Copyright © 2013 Wiley Periodicals, Inc.)

Background: MicroRNAs (miRNAs) are small RNAs that regulate gene expression by suppressing protein translation and may influence RNA expression. MicroRNAs are detected in extracellular locations such as plasma; however, the extent of miRNA expression in plasma its relation to cardiovascular disease is not clear and many clinical studies have utilized array-based platforms with poor reproducibility., Methods and Results: Initially, to define distribution of miRNA in human blood; whole blood, platelets, mononuclear cells, plasma, and serum from 5 normal individuals were screened for 852 miRNAs using high-throughput micro-fluidic quantitative RT-PCR (qRT-PCR). In total; 609, 448, 658, 147, and 178 miRNAs were found to be expressed in moderate to high levels in whole blood, platelets, mononuclear cells, plasma, and serum, respectively, with some miRNAs uniquely expressed. To determine the cardiovascular relevance of blood miRNA expression, plasma miRNA (n=852) levels were measured in 83 patients presenting for cardiac catheterization. Eight plasma miRNAs were found to have over 2-fold increased expression in patients with significant coronary disease (≥70% stenosis) as compared to those with minimal coronary disease (less than 70% stenosis) or normal coronary arteries. Expression of miR-494, miR-490-3p, and miR-769-3p were found to have significantly different levels of expression. Using a multivariable regression model including cardiovascular risk factors and medications, hsa-miR-769-3p was found to be significantly correlated with the presence of significant coronary atherosclerosis., Conclusions: This study utilized a superior high-throughput qRT-PCR based method and found that miRNAs are found to be widely expressed in human blood with differences expressed between cellular and extracellular fractions. Importantly, specific miRNAs from circulating plasma are associated with the presence of significant coronary disease.

Objective: The purpose of this study was to characterize the relationship between adipose tissue phenotype and depot-specific microvascular function in fat., Methods and Results: In 30 obese subjects (age 42±11 years, body mass index 46±11 kg/m(2)) undergoing bariatric surgery, we intraoperatively collected visceral and subcutaneous adipose tissue and characterized depot-specific adipose phenotypes. We assessed vasomotor function of the adipose microvasculature using videomicroscopy of small arterioles (75-250 μm) isolated from different fat compartments. Endothelium-dependent, acetylcholine-mediated vasodilation was severely impaired in visceral arterioles, compared to the subcutaneous depot (P<0.001 by ANOVA). Nonendothelium dependent responses to papaverine and nitroprusside were similar. Endothelial nitric oxide synthase inhibition with N(ω)-nitro-l-arginine methyl ester reduced subcutaneous vasodilation but had no effect on severely blunted visceral arteriolar responses. Visceral fat exhibited greater expression of proinflammatory, oxidative stress-related, hypoxia-induced, and proangiogenic genes; increased activated macrophage populations; and had a higher capacity for cytokine production ex vivo., Conclusions: Our findings provide clinical evidence that the visceral microenvironment may be intrinsically toxic to arterial health providing a potential mechanism by which visceral adiposity burden is linked to atherosclerotic vascular disease. Our findings also support the evolving concept that both adipose tissue quality and quantity may play significant roles in shaping cardiovascular phenotypes in human obesity.

A significant need exists for orthopedic implants that can intrinsically resist bacterial colonization. In this study, three biomaterials that are used in spinal implants--titanium (Ti), polyether-ether-ketone (PEEK), and silicon nitride (Si₃N₄)--were tested to understand their respective susceptibility to bacterial infection with Staphylococcus epidermidis, Staphlococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Enterococcus. Specifically, the surface chemistry, wettability, and nanostructured topography of respective biomaterials, and the effects on bacterial biofilm formation, colonization, and growth were investigated. Ti and PEEK were received with as-machined surfaces; both materials are hydrophobic, with net negative surface charges. Two surface finishes of Si₃N₄ were examined: as-fired and polished. In contrast to Ti and PEEK, the surface of Si₃N₄ is hydrophilic, with a net positive charge. A decreased biofilm formation was found, as well as fewer live bacteria on both the as-fired and polished Si₃N₄. These differences may reflect differential surface chemistry and surface nanostructure properties between the biomaterials tested. Because protein adsorption on material surfaces affects bacterial adhesion, the adsorption of fibronectin, vitronectin, and laminin on Ti, PEEK, and Si₃N₄ were also examined. Significantly greater amounts of these proteins adhered to Si₃N₄ than to Ti or PEEK. The findings of this study suggest that surface properties of biomaterials lead to differential adsorption of physiologic proteins, and that this phenomenon could explain the observed in-vitro differences in bacterial affinity for the respective biomaterials. Intrinsic biomaterial properties as they relate to resistance to bacterial colonization may reflect a novel strategy toward designing future orthopedic implants.

Bacterial infection of in-dwelling medical devices is a growing problem that cannot be treated by traditional antibiotics due to the increasing prevalence of antimicrobial resistance and biofilm formation. Here, due to changes in surface parameters, it is proposed that bacterial adhesion can be prevented through nanosurface modifications of the medical device alone. Toward this goal, titanium was created to possess nanotubular surface topographies of highly controlled diameters of 20, 40, 60, or 80 nm, sometimes followed by heat treatment to control chemistry and crystallinity, through a novel anodization process. For the first time it was found that through the control of Ti surface parameters including chemistry, crystallinity, nanotube size, and hydrophilicity, significantly changed responses of both Staphylococcus epidermidis and Staphylococcus aureus (pathogens relevant for orthopaedic and other medical device related infections) were measured. Specifically, heat treatment of 80 nm diameter titanium tubes produced the most robust antimicrobial effect of all surface treatment parameters tested. This study provides the first step toward understanding the surface properties of nano-structured titanium that improve tissue growth (as has been previously observed with nanotubular titanium), while simultaneously reducing infection without the use of pharmaceutical drugs.

Infection is a major problem in orthopedics leading to implant failure. It is a challenging task to treat orthopedic implant infection, which may lead to implant replacement and, in severe cases, may result in amputation and mortality. Infection poses an even further risk as bacteria are beginning to develop resistance against commonly used antibiotics. Therefore, in this research a combination of various approaches was used to fight implant infection without resorting to the use of antibiotics. Specifically, conventional titanium was altered through a process of anodization and electrical stimulation to reduce Staphylococcus aureus growth. It was shown that when a 15-30 V electrical stimulation was coupled with anodized nanotubular titanium a significant decrease in S. aureus biofilm formation was observed, compared with non-anodized and non-electrically stimulated titanium after 2 days culture. The decrease in biofilm formation observed here was explained by the presence of fluorine on the surfaces of anodized nanotubular titanium. Thus, coupling the positive influences of anodization and electrical stimulation could be a promising way to fight titanium-based orthopedic device-related infections., (Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

One of the important prerequisites for a successful orthopedic implant apart from being osteoconductive is the elicitation of a favorable immune response that does not lead to the rejection of the implant by the host tissue. Anodization is one of the simplest surface modification processes used to create nanotextured and nanotubular features on metal oxides which has been shown to improve bone formation. Anodization of titanium (Ti) leads to the formation of TiO(2) nanotubes on the surface, and the presence of these nanotubes mimics the natural nanoscale features of bone, which in turn contributes to improved bone cell attachment, migration, and proliferation. However, inflammatory cell responses on anodized Ti remains to be tested. It is hypothesized that surface roughness and surface feature size on anodized Ti can be carefully manipulated to control immune cell (specifically, macrophages) responses. Here, when Ti samples were anodized at 10 V in the presence of 1% hydrofluoric acid (HF) for 1 minute, nanotextured (nonnanotube) surfaces were created. When anodization of Ti samples was carried out with 1% HF for 10 minutes at 15 V, nanotubes with 40-50 nm diameters were formed, whereas at 20 V with 1% HF for 10 minutes, nanotubes with 60-70 nm diameters were formed. In this study, a reduced density of macrophages was observed after 24 hours of culture on nanotextured and nanotubular Ti samples which were anodized at 10, 15, and 20 V, compared with conventional unmodified Ti samples. This in vitro study thus demonstrated a reduced density of macrophages on anodized Ti, thereby providing further evidence of the greater efficacy of anodized Ti for orthopedic applications.

Materials currently used for bladder applications often suffer from incomplete coverage by urothelial cells (cells that line the interior of the bladder and ureter) which leads to the continuous exposure of the underlying materials aggravating an immune response. In particular, a ureteral (or sometimes called an ureteric or bladder) stent is a thin tube inserted into the ureter to prevent or treat obstruction of urine flow from the kidney. The main complications with ureteral stents are infection and blockage by encrustation, which can be avoided by promoting the formation of a monolayer of urothelial cells on the surface of the stent. Nanotechnology (or the use of nanomaterials) may aid in urothelialization of bladder stents since nanomaterials have been shown to have unique surface energetics to promote the adsorption of proteins important for urothelial cell adhesion and proliferation. Since many bladder stents are composed of titanium, this study investigated the attachment and spreading of human urothelial cells on different nanotextured titanium surfaces. An inexpensive and effective scaled up anodization process was used to create equally distributed nanotubular surfaces of different diameter sizes from 20-80 nm on titanium with lengths approximately 500 nm. Results showed that compared to untreated titanium stents and 80 nm diameter nanotubular titanium, 20 nm diameter nanotubular titanium stents enhanced human urothelial cell adhesion and growth up to 3 days in culture. In this manner, this study suggests that titanium anodized to possess nanotubular surface features should be further explored for bladder stent applications.

Pure as well as Mg(2+)- and F(-)-doped nanophase (i.e., grain sizes in the nanometer regime in at least one dimension) hydroxyapatite (HA) samples were synthesized by a precipitation method followed by sintering at 1100 degrees C for 1 h to determine their microstructural, mechanical, and osteoblast (bone-forming cell) adhesion properties pertinent for orthopedic applications. Different amounts of Mg(2+) and F(-) ions (specifically from 0 to 7.5 mol %) were doped into the HA samples. X-ray diffraction was used to identify the presence of crystalline phases, lattice parameters, and crystal volumes of the samples. Fourier transform infrared (FTIR) was further used to chemically characterize HA, and thus FTIR patterns revealed the characteristic absorption bands of HA. Microhardness measurements were also performed to assess mechanical properties of the novel formulations. Results of this study showed an improvement in sample density for some of the samples, which was a consequence of the molar percentage variation of the dopants. Moreover, in most of the samples doped with Mg, beta-tricalcium phosphate was observed as a second phase to HA. In addition, 1% Mg- and 2.5% F-doped HA had the highest microhardness values. Lastly, results demonstrated the highest osteoblast densities when the HA samples were doped with 2.5-7.5% Mg(2+) and F(-). Thus, the results of this study suggest that decreasing the grain size of HA into the nanometer regime and doping HA with Mg(2+) and F(-) can potentially increase the efficacy of HA for orthopedic applications., ((c) 2010 Wiley Periodicals, Inc.)

Over the past decade, nanotechnology (or the use of materials with dimensions less than 100 nm in at least one direction) has been proposed to improve the lifespan of many biomedical devices, including orthopedic implants. Specifically, to improve the cytocompatibility properties of currently used orthopedic implants, nanotechnology has been used to create nanometer surface features (through anodization) on titanium. In addition to this approach, another therapeutic method widely investigated to heal bone fractures is through electrical stimulation. Here, the coupling of such nanotechnology approaches and electrical stimulation were studied to maximize bone cell functions on titanium. Results showed that compared to unstimulated conventional titanium, bone forming cell (osteoblast) proliferation and long-term functions (alkaline phosphatase synthesis, collagen type I synthesis and calcium deposition) were improved upon both the creation of an anodized nanotubular titanium surface and biphasic electrical stimulation. Most importantly, when electrical stimulation was combined with anodized nanotubular titanium features, osteoblast long-term functions were improved the most. Therefore, coupling the positive effects of anodized nanotubular titanium topographies with currently used therapeutic electrical stimulation should be further studied to improve orthopedic implants., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Zirconia (ZrO(2)) and barium sulfate (BaSO(4)) particles were introduced into a methyl methacrylate monomer (MMA) solution with polymethyl methacrylate (PMMA) beads during polymerization to develop the following novel bone cements: bone cements with unfunctionalized ZrO(2) micron particles, bone cements with unfunctionalized ZrO(2) nanoparticles, bone cements with ZrO(2) nanoparticles functionalized with 3-(trimethoxysilyl)propyl methacrylate (TMS), bone cements with unfunctionalized BaSO(4) micron particles, bone cements with unfunctionalized BaSO(4) nanoparticles, and bone cements with BaSO(4) nanoparticles functionalized with TMS. Results demonstrated that in vitro osteoblast (bone-forming cell) densities were greater on bone cements containing BaSO(4) ceramic particles after four hours compared to control unmodified bone cements. Osteoblast densities were also greater on bone cements containing all of the ceramic particles after 24 hours compared to unmodified bone cements, particularly those bone cements containing nanofunctionalized ceramic particles. Bone cements containing ceramic particles demonstrated significantly altered mechanical properties; specifically, under tensile loading, plain bone cements and bone cements containing unfunctionalized ceramic particles exhibited brittle failure modes whereas bone cements containing nanofunctionalized ceramic particles exhibited plastic failure modes. Finally, all bone cements containing ceramic particles possessed greater radio-opacity than unmodified bone cements. In summary, the results of this study demonstrated a positive impact on the properties of traditional bone cements for orthopedic applications with the addition of unfunctionalized and TMS functionalized ceramic nanoparticles.

Purpose: The precise molecular mechanisms culminating in coronary artery disease (CAD) are not well understood, despite a wealth of knowledge on predisposing risk factors and pathomechanisms. CAD and myocardial infarction (MI) are complex genetic diseases; neither the environment alone, nor a single gene, cause disease, rather, a mix of environmental and genetic factors lead to atherosclerosis of the coronary arteries., Materials and Methods: In the present study, our aim was to investigate the roles of prothrombin G20210A mutation and Factor VLeiden mutation in atherosclerotic coronary artery disease. 287 subjects (106 control subjects, who were angiographically normal, and 181 angiographically documented coronary atherosclerotic patients who exhibited coronary artery narrowing to a degree of >or=50%) were included in this study. The mutations were assessed with LightCycler Real-Time PCR mutation detection kits (Roche Diagnostics, GmbH, Germany)., Results: 6.6% of control subjects, and 6.1% of patients with (50% coronary artery narrowing were determined to have the Factor VLeiden heterozygote mutation. 6.6% of control subjects had the Prothrombin G20210A heterozygote mutation, while 7.7% of patients with (50% coronary artery narrowing had this mutation. The OR for Factor VLeiden was 1.52 (CI: 0.240-9.602) and for Prothrombin G20210A mutation, the OR was 1.415 (CI: 0.287-6.962)., Conclusion: Although both the heterozygote Factor VLeiden and Prothrombin gene mutations were more frequent in patients with CAD than in control subjects, there was no statistical relationship found to exist between coronary artery disease and the Factor VLeiden and Prothrombin G20210A mutations.

A new method for the extraction of both anions and cations is proposed using electro-synthesized polypyrrole (PPy) and overoxidized sulfonated polypyrrole film (OSPPy). In situ anion (chloride, nitrate, sulfate) and cation (calcium, magnesium) uptake and release were examined under controlled potential conditions for prospective applications in electrochemically controlled solid-phase extraction (EC-SPE). The PPy film was used as an anode (anion-exchanger) and OSPPy film was used as a cathode (cation-exchanger) material and reverse order of the electrodes were investigated in EC-SPE. This new cell arrangement containing two ion exchanger polymer electrodes was developed to provide in situ removal of both anions and cations from aqueous solution. Simple preparation of the film coatings on a platinum plate was possible using a constant potential method. Applied positive and negative potentials facilitated the in situ extraction and desorption of ions, respectively. Both anions and cations were desorbed into sample aliquot and were determined by ion chromatography (IC). The method was validated using a standard reference material and tested for the determination of the ions in real water samples.

Pancreatic cancer has been linked with exposure to environmental chemicals, which generally require metabolic activation to highly reactive toxic or carcinogenic intermediates. N-acetyltransferase 1 (NAT1) and N-acetyltransferase 2 (NAT2) are expressed primarily in extrahepatic and hepatic tissues, respectively. Both enzymes catalyze N- and O-acetylation of aromatic and heterocyclic amines. It is believed that these compounds are activated via O-acetylation and detoxified by N-acetylation. Several polymorphisms of these two genes have been associated with an increased risk of cancer. Twenty-seven cases of pancreatic cancer and 104 controls were included in this study. Blood was collected in EDTA-containing tubes, and genomic DNA was extracted from the white blood cells by using a high pure PCR template preparation kit. Genotyping of NAT2 polymorphisms was detected by a real time PCR instrument. There was a significant difference in the distribution of the NAT2*6A acetylators phenotype between cases and the controls. The odds ratio of pancreatic cancer for the NAT2*6A slow phenotype was 5.7 (95% CI = 1.27-25.55; p = 0.023) compared with the fast type. Our results suggest that slow acetylators have higher risk of developing pancreatic cancer than fast acetylators. NAT2 gene polymorphisms may be associated with genetic susceptibility to pancreatic cancer.

Currently used orthopedic implants composed of titanium have a limited functional lifetime of only 10-15 years. One of the reasons for this persistent problem is the poor prolonged ability of titanium to remain bonded to juxtaposed bone. It has been proposed to modify titanium through anodization to create a novel nanotubular topography in order to improve cytocompatibility properties necessary for the prolonged attachment of orthopedic implants to surrounding bone. Additionally, electrical stimulation has been used in orthopedics to heal bone non-unions and fractures in anatomically difficult to operate sites (such as the spine). In this study, these two approaches were combined as the efficacy of electrical stimulation to promote osteoblast (bone forming cell) density on anodized titanium was investigated. To do this, osteoblast proliferation experiments lasting up to 5 days were conducted as cells were stimulated with constant bipolar pulses at a frequency of 20 Hz and a pulse duration of 0.4 ms each day for 1 hour. The stimulation voltages were 1 V, 5 V, 10 V, and 15 V. Results showed for the first time that under electrical stimulation, osteoblast proliferation on anodized titanium was enhanced at lower voltages compared to what was observed on conventional (nonanodized) titanium. In addition, compared to nonstimulated conventional titanium, osteoblast proliferation was enhanced 72% after 5 days of culture on anodized nanotubular titanium at 15 V of electrical stimulation. Thus, results of this study suggest that coupling the positive influences of electrical stimulation and nanotubular features on anodized titanium may improve osteoblast responses necessary for enhanced orthopedic implant efficacy.

Advancements in nanobiotechnology are revolutionizing our capability to understand biological intricacies and resolve biological and medical problems by developing subtle biomimetic techniques. Nanocomposites and nanostructured materials are believed to play a pivotal role in orthopedic research since bone itself is a typical example of a nanocomposite. This article reviews current strategies using nanobiomaterials to improve current orthopedic materials and examines their applications in bone tissue engineering. Preliminary investigations support the potential of nanobiomaterials in orthopedic applications; however, significant advancements are necessary to achieve clinical use. Overall, current trends in nanobiotechnology foreshadow a bright future through the use of nanobiomaterials in the orthopedic domain., ((c) 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.)

Although smoking is regarded as the most important causal factor in chronic obstructive pulmonary disease (COPD), only 10-20% of smokers develop symptomatic COPD, which indicates the presence of genetic predisposing factors in its pathogenesis. This study investigates the association between gene polymorphysims of glutathione S-transferases (GSTs) and COPD. Blood samples were taken from 149 patients and 150 healthy controls. Polymorphisms of GSTT1, GSTM1, and GSTP1 were genotyped using Real-Time PCR. Multivariate logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals between specific genotypes and COPD. There was no difference in the frequencies of the genotypes of GSTM1 and GSTT1 between the groups, but the GSTP1 Ile/Ile genotype was significantly higher in the patients than in the controls (61.1% vs. 38%). GSTP1 Ile/Val and Val/Val genotypes were associated with a decreased risk of COPD when compared to the Ile/Ile genotype (2.12-fold and 4-fold, respectively). Thus we suggest that the Val allele of GSTP1 may have a protective effect for development of COPD. Furthermore, when we evaluated the association between GSTP1 genes and smoking status, smokers with the GSTP1 Ile allele had an increased risk for the development of COPD. Among the combinations of the genotypes, the combination of GSTM1, GSTT1 null, and GSTP1 Val/Val was associated with the maximal increased risk (12-fold) of COPD. Thus to explain the ethiopathogenesis of COPD, investigation of a single gene family is inadequate. Based on our results and the previous data, further studies should be focused on the GSTP1 gene and the interactions with other genes such as polymorphisms of N-acetyltransferases, GSTM1 and GSTT1, microsomal epoxide hydrolase, and allelic variants of cytochrome P450.

In obstructive jaundice, free radical production is increased and antioxidative activity is reduced. N-Acetylcysteine (NAC) has a beneficial effect with anti-inflammatory and antioxidant activity, acting as a free radical scavenger. NAC inhibits inducible nitric oxide synthase, suppresses cytokine expression/release, and inhibits adhesion molecule expression and nuclear factor kappa B. The aim of this study was to investigate the effects of NAC on liver/renal tissue and serum lipid peroxidation in lipopolysaccharide (LPS)-induced obstructive jaundice. We randomized 60 rats into 6 groups: group 1, Sham; group 2, obstructive jaundice (OJ) induced after bile-duct ligation; group 3, OJ + NAC (100 mg kg- 1 subcutaneously); group 4, OJ + LPS (10 mg kg-1); group 5, OJ + NAC + LPS; and group 6, OJ + LPS + NAC. For each group, the biochemical markers of lipid peroxidation and the antioxidant products were measured in serum and liver/renal tissue after sacrifice. Almost all lipid peroxidation products levels were increased and antioxidant products levels were decreased in groups who received LPS (groups 4, 5, and 6), but the effect was less remarkable when NAC was administered before LPS (group 5). The same trend was seen for groups with OJ +/- LPS who did not received NAC or received it after induced toxemia (groups 2, 4, and 6) as compared to groups 1 and 3. Moreover, in the case of OJ + LPS, rats treated with NAC before LPS (group 5) had lower lipid peroxidation products levels and higher antioxidant products levels as compared to those who did not received NAC (group 4). This phenomenon was not reproducible with NAC administered after LPS (group 6). Thus, results of this study showed that NAC prevents the deleterious effects of LPS in obstructive jaundice by reducing lipid peroxidation in serum and liver/renal tissue if administered before LPS. Nonetheless, NAC failed to prevent the lipid peroxidation in the case of established endotoxemia in obstructive jaundice.

The acetylation polymorphism is a common inherited variation in human drug and carcinogen metabolism. Because N- acetyltransferase (NAT2) is important for the detoxification and/or bioactivation of drugs and carcinogens, polymorphisms of this gene have important implications in therapeutics of and susceptibility to cancer. In this study, NAT2 genotype (NAT2*5A (C(481)T), NAT2*6A (G(590)A), NAT2*7A/B (G(857)A)) and NAT2*14A (G(191)A) and phenotype were determined in 125 patients with colorectal carcinoma and 82 healthy control in Mersin, a city located in the southern region of Turkey. Isolation of the subjects' DNA was performed by using a highly purified PCR template preparation kit/(Roche Diagnostics cat. no: 1 796 828) and the NAT2 polymorphism was detected using real-time PCR (Roche Diagnostics, GmbH, Mannheim, Germany). According to this study high protein intake is associated with the increased risk for the development of colon cancer (OR = 1.73; 95% CI, 1.10-3.07). Although only NAT2*14A fast type was associated with increased risk in patients with colorectal carcinoma (OR = 3.03; 95% CI, 1.56-5.86), when a high protein diet was considered, NAT2*7A/B fast genotype was also found to be associated with an increased risk (OR = 2.06, 95% CI for NAT2*7A/B, 1.10-3.86; OR = 2.65; 95% CI, 1.29-5.46 for NAT2*14A). Smoking status did not differ between the control and patient groups. Our data suggest that exposure to carcinogens through consumption of a high-protein diet may increase the risk of colorectal carcinoma only in genetically-susceptible individuals., (Copyright 2005 John Wiley & Sons, Ltd.)

Objective: The exact mechanism of the increased cardiovascular morbidity and mortality in type-2 diabetes is still undefined. The aim of our study was to assess the impact of apolipoprotein E (apo E) polymorphism and other factors on atherosclerotic vascular disease in type-2 diabetic patients. We also examined the association between apo E polymorphism and lipid profile in diabetic patients., Methods and Results: We assessed the apo E polymorphism in 295 atherosclerotic patients (124 of them had diabetes (according to WHO criteria) and 171 of them had coronary artery narrowing > 50). The detection of apo E polymorphism was made by Real-Time PCR using a Light-Cycler (Roche diagnostics, GmbH, Mannheim, Germany). Serum triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), lipoprotein (a) [Lp(a)], apolipoprotein A (Apo A) and apolipoprotein B (Apo B) levels were determined by biochemical analyser. Genotypic distribution of apo E polymorphism did not differ between diabetic and non-diabetic atherosclerotic patients. The distributions of apo E2/2, E2/3, E2/4, E3/3, E3/4 and E4/4 genotypes in diabetic and non-diabetic atherosclerotic patients were 7.3%: 8.2%, 15.3%: 15.8%, 4.0%: 5.3%, 50.8%: 56.7%, 16.9%: 11.1% and 5.6%: 2.9%, respectively. Participants were grouped as apo E2 (E2/2 or E2/3), apo E3 (E3/3), or apo E4 (E4/4 or E4/3). The distributions of apo E2, E3 and E4 alleles were 23.5%, 52.9%, 23.5%, for diabetic patients, and 25.3%, 59.9%, 14.8% for non-diabetic patients, respectively. The apolipoprotein E genotype was not associated with the lipid levels in diabetic patients., Conclusions: Our findings imply that apo E polymorphism is not related to the development of atherosclerosis in patients with type-2 diabetes.

Glutathione S-transferases (GSTs) belong to a superfamily of detoxification enzymes that provide critical defences against a large variety of chemical carcinogens and environmental toxicants. GSTs are present in most epithelial tissues of the human gastrointestinal tract. We investigated associations between genetic variability in specific GST genes (GSTM1, GSTT1 and GSTP1), the interaction with cigarette smoking and susceptibility to gastric cancer. The GSTM1, GSTT1 and GSTP1 polymorphisms were determined using real-time polymerase chain reaction (PCR) and fluorescence resonance energy transfer with Light Cycler Instrument. The study included 70 patients with gastric cancer and 204 controls. Associations between specific genotypes and the development of gastric cancer were examined by use of logistic regression to calculate odds ratios (OR) and 95% confidence intervals (CI). The GSTM1 homozygous null genotype was associated with an increased risk of developing gastric cancer (OR = 1.73; 95% CI = 1.10-3.04). GSTT1 homozygous null genotype and GSTP1 genotypes were not associated with the risk of gastric cancer. Also there was no difference between cases and controls in the frequency of val-105 and ile-105 alleles (p = 0.07). After grouping according to smoking status, GSTM1 null genotype was associated with an increased gastric cancer risk for smokers (OR = 2.15; 95% CI, 1.02-4.52). There were no significant differences in the distributions of any of the other GST gene combinations. Our findings suggest that the GSTM1 null genotype may be associated with an increased susceptibility to gastric cancer., (Copyright 2004 John Wiley & Sons, Ltd.)

In the present study, Nigella unguicularis (Lam.) Spenner (Ranunculaceae) (Nu) fixed oil was administered orally to Wistar Kyoto rats for 4 weeks. The effects of the oil on serum lipid profile, haematological parameters and oxidant/antioxidant balance were investigated. The study showed that daily administration of the oil (1 ml/kg orally for 4 weeks) caused a significant decrease in serum total cholesterol, VLDL-cholesterol, triglycerides, glucose levels and a significant elevation of serum high-density lipoprotein level. The serum transaminases ( aspartate transaminase (AST), alanine transaminase (ALT)), alkaline phosphatase, bilirubin, blood urea nitrogen, urea, mean corpuscular haemoglobin concentration decreased significantly while albumin, mean corpuscular volume and fibrinogen levels increased significantly compared to control values. The administration of the oil (1 ml/kg orally for 4 weeks) caused a significant increase total antioxidant status in rats. Treatment with Nigella unguicularis oil did not effect malondialdehyde (MDA) concentrations. It is concluded that Nigella unguicularis oil possesses favourable metabolic effects on blood biochemistry and oxidant/antioxidant balance in rats.

Hypothesis: Glutathione and glutathione-related antioxidant enzymes are involved in the metabolism and detoxification of cytotoxic and carcinogenic compounds as well as reactive oxygen species. Reactive oxygen species generation occurs in prolonged relative hypoperfusion conditions such as in aging. The etiology of presbycusis is much less certain; however, a complex genetic cause is most likely. The effect of aging shows a wide interindividual range; we aimed to investigate whether profiles of (glutathione S-transferase (GST) M1, T1 and P1 genotypes may be associated with the risk of age-related hearing loss., Patients and Methods: We examined 68 adults with presbycusis and 69 healthy controls. DNA was extracted from whole blood, and the GSTM1, GSTT1 and GSTP1 polymorphisms were determined using a real-time polymerase chain reaction and fluorescence resonance energy transfer with a Light-Cycler Instrument. Associations between specific genotypes and the development of presbycusis were examined by use of logistic regression analyses to calculate odds ratios and 95% confidence intervals., Results: Gene polymorphisms at GSTM1, GSTT1, and GSTP1 in subjects with presbycusis were not significantly different than in the controls (p > 0.05). Also, the combinations of different GSTM1, GSTT1, and GSTP1 genotypes were not an increased risk of presbycusis (p > 0.05)., Conclusion: We could not demonstrate any significant association between the GSTM1, GSTT1, and GSTP1 polymorphism and age-related hearing loss in this population. This may be because of our sample size, and further studies need to investigate the exact role of GST gene polymorphisms in the etiopathogenesis of the presbycusis.

The role of free radicals in fibromyalgia is controversial. In this study, 85 female patients with primary fibromyalgia and 80 age-, height-, and weight-matched healthy women were evaluated for oxidant/antioxidant balance. Malondialdehyde is a toxic metabolite of lipid peroxidation used as a marker of free radical damage. Superoxide dismutase is an intracellular antioxidant enzyme and shows antioxidant capacity. Pain was assessed by visual analog scale. Tender points were assessed by palpation. Age, smoking, body mass index (BMI), and duration of disease were also recorded. Malondialdehyde levels were significantly higher and superoxide dismutase levels significantly lower in fibromyalgic patients than controls. Age, BMI, smoking, and duration of disease did not affect these parameters. We found no correlation between pain and number of tender points. In conclusion, oxidant/antioxidant balances were changed in fibromyalgia. Increased free radical levels may be responsible for the development of fibromyalgia. These findings may support the hypothesis of fibromyalgia as an oxidative disorder.

The glutathione S-transferase (GST) supergene family is an important part of cellular enzyme defense against endogenous and exogenous chemicals, many of which have carcinogenic potential. The present investigation was conducted to detect a possible association between polymorphisms at the GSTM1, GSTT1, and GSTP1 genes and the interaction with cigarette smoking and colorectal cancer incidence. We examined 181 patients with colorectal cancer and 204 controls. DNA was extracted from whole blood, and the GSTM1, GSTT1, and GSTP1 polymorphisms were determined using a real-time polymerase chain reaction and fluorescence resonance energy transfer with a Light-Cycler instrument. Associations between specific genotypes and the development of colorectal cancer were examined by use of logistic regression analysis to calculate odds ratios (OR) and 95% confidence intervals (CI). The GSTM1 polymorphism was associated with an increased risk of developing colorectal cancer (OR = 1.62, 95% CI: 1.06-2.46). Also the risk of colorectal cancer associated with the GSTT1 null genotype was 1.64 (95% CI: 1.10-2.59). Statistically no differences were found between patients with colorectal cancer and control groups for the GSTP1 Ile/Ile, Ile/Val and Val/Val genotypes. In addition, the frequencies of the GSTM1 and GSTT1 deletion genotypes differed significantly between the cases and controls for current smokers; the GSTT1 null genotype especially is associated with a greater risk of colorectal cancer (OR = 2.44, 95% CI: 1.24-4.81). The GSTM1 and GSTT1 deletions were associated with an increased risk of developing a transverse or rectal tumor (OR = 1.86, 95% CI: 1.15-3.00; OR = 1.70, 95% CI: 1.02-2.84; respectively). The glutathione S-transferase polymorphisms were not associated with risk in patients stratified by age. The risk of colorectal cancer increased as putative high-risk genotypes increased for the combined genotypes of GSTM1 null, GSTT1 null, and either GSTP1 valine heterozygosity or GSTP1 valine homozygosity (OR = 2.69, 95% CI: 1.02-7.11). In conclusion, the results obtained in this study clearly suggest that those susceptibility factors related to different GST polymorphic enzymes are predisposing for colorectal cancer.

Background: To evaluate the relationship between the serum oxidant-antioxidant balance and the presence of exfoliation syndrome (XFS) in a prospective study., Methods: The study group comprised 27 cases of XFS. Serum concentrations of myeloperoxidase, malondialdehyde, vitamins A, C and E, catalase and total antioxidant were measured and compared with an independent group by t-test., Results: Serum myeloperoxidase, vitamins A and E, catalase and total antioxidant parameters were not statistically different among the groups (P >0.05). Serum vitamin C concentrations were significantly lower and malondialdehyde concentrations were significantly higher in cases of XFS when compared with the control group (P=0.008 and P=0.002, respectively)., Conclusion: An altered oxidant-antioxidant balance may play a role in the pathogenesis of XFS.

Objectives: Asthma is a complex multifactorial disease with an obvious genetic predisposition, immunological aberration, and involvement of noxious environmental factors. Polymorphisms of the glutathione-S-transferase (GST) genes are known risk factors for some environmentally-related diseases. In the present study, the hypothesis that polymorphisms in the GSTT1, GSTM1 and GSTP1 genes are associated with atopic and nonatopic asthma was examined., Methodology: The study population consisted of 103 unrelated healthy individuals and 101 patients with bronchial asthma (64 atopic, 37 nonatopic). Asthma was diagnosed according to the American Thoracic Society statement. Genotyping of polymorphisms in the GSTT1, GSTM1 and GSTP1 genes was performed using real time polymerase chain reaction with a Light Cycler instrument and hybridization probes in combination with the Light Cycler DNA master hybridization probes kit., Results: Patients with atopic asthma (34.4%) had a higher prevalence of the GSTT1 null genotype than the nonatopic asthma patients (13.5%; OR = 3.83; 95% CI, 1.24-11.78). Asthma patients (63.4%) had a higher prevalence of the GSTM1 null genotype than the control group (40.8%; OR = 2.34; 95% CI, 1.31-4.20). Subjects with the GSTP1 homozygous Val/Val genotype had a 3.55-fold increased risk of having atopic asthma compared to nonatopic asthma (OR = 3.55; 95% CI, 1.10-12.56)., Conclusions: These results suggest that the GSTT1 and GSTM1 null genotypes and the GSTP1 Val/Val polymorphism may play important roles in asthma pathogenesis. It is possible that intermediate electrophilic metabolites, arising in the first phase of detoxification, are not metabolized by GST enzymes in asthmatic patients and are not excreted. These intermediate metabolites may damage cells and generate oxidative stress, and so contribute to the pathogenesis of asthma.

The antioxidant and anti-inflammatory properties of N-acetylcysteine has been documented in many experimental lung injury models. Because intravenous injection of oleic acid induces histopathologic changes similar to those seen in human acute lung injury or acute respiratory distress syndrome, the authors evaluated the effects of N-acetylcysteine (NAC) on oxidative stress and lung damage in an oleic acid (OA)-induced lung injury model. Thirty-five rats were divided into 5 groups as sham, NAC, OA, pre-OA-NAC, and post-OA-NAC. Lung damage was induced by intravenous administration of oleic acid. Pre-OA-NACgroup received intravenous (IV) N-acetylcysteine 15 minutes before oleic acid infusion and post-OA-NAC group received IV N-acetylcysteine 2 hours after oleic acid infusion. In both of the N-acetylcysteine treatment groups, blood and tissue samples were collected 4 hours after oleic acid infusion, independent from the time of N-acetylcysteine infusion. In other groups, blood and tissue samples were collected 4 hours after ethanol, NAC, or OA infusions. Serum myeloperoxidase activity, total antioxidant capacity, malondialdehyde levels, and lung tissue Na+ - K+ ATPase activity were measured and light microscopic analyses of lung specimens were performed. The administration of N-acetylcysteine significantly restored Na+ - K+ ATPase activity and total antioxidant capacity levels and ameliorated lung architecture. N-acetylcysteine has been shown to have some attenuating effects in experimental animal studies. However, further investigations are necessary to suggest N-acetylcysteine as a treatment agent in critically ill patients with lung injury.

Reactive oxygen species have been implicated in pathogenesis injury after ischaemia-reperfusion (I/R). Caffeic Acid Phenethyl Ester (CAPE), an active component of honeybee propolis extract, exhibits antioxidant and anti-inflammatory properties. The aim of this study was to investigate the effects of CAPE on erythrocyte membrane damage after hind limb I/R. Rats were divided into two groups: I/R and I/R with CAPE pre-treatment. They were anaesthetized with intramuscular ketamine 100 mg kg(-1). A 4-h I/R period was performed on the right hind limb of all animals. In the CAPE-treated group, animals received CAPE 10 microm by intraperitoneal injection 1 h before the reperfusion. At the end of the reperfusion period, a midsternotomy was performed. A 5-ml blood sample was withdrawn from the ascending aorta for biochemical assays. Serum and erythrocyte membrane MDA levels were significantly lower in the CAPE-treated group when compared to the I/R group ( p = 0.001 and p<0.001, respectively). Erythrocyte membrane Na(+)-K(+) ATPases activity in the CAPE-treated group was significantly higher than the I/R group ( p<0.001). In conclusion, CAPE seems to be effective in protecting against oxidative stress. Therefore, we suggest that in order to decrease I/R injury, pre-administration of CAPE may be a promising agent for a variety of conditions associated with I/R.

Purpose: To investigate the influence of the glutathione S-transferase (GST) M1, T1, and P1 genotypes on the laryngeal squamous cell carcinoma risk., Material and Methods: The study group consisted of 42 white patients with laryngeal squamous cell carcinoma (39 of them were male, mean age: 53, range: 37-67 and 3 of them were female, mean age: 47, range: 32-55) and 89 control subjects (nonsmokers = 47, smokers = 42) (58 male and 31 female, mean age: 51, range: 30-72). DNA samples were isolated from blood samples using high pure polymerase chain reaction (PCR) Template Preparation Kit. The detection of GST T1, GST M1, and P1 polymorphisms were detected by using real-time PCR., Results: Gene polymorphisms at GST M1 and P1 were not significantly different in patient and control groups. However, GST T1 null type significantly increased in laryngeal cancer patients when compared with the nonsmoking controls (P =.04)., Conclusions: There was a significant association between GST T1 null genotype and laryngeal squamous cell carcinoma. However the potential role of GSTs as markers of susceptibility to laryngeal carcinoma needs further studies in a larger number of patients.

Coronary artery disease (CAD) is the leading cause of morbidity and mortality in the world, and cigarette smoking is a major contributing factor to the disease. Glutathione S-transferase (GST) enzyme is implicated in the detoxification of carcinogens present in tobacco smoke and consequent polymorphisms in this gene may confer susceptibility to cardiovascular disease if DNA damage is important in CAD. Therefore, we examined this question in a case-control study of subjects having coronary atheroma by angiography and with a past history of myocardial infarction (MI). The study population consists of 247 healthy controls and 148 consecutive patients who had undergone coronary angiography for suspicion of coronary artery disease. DNA was extracted from whole blood, and the GSTM1 and GSTT1 polymorphisms were determined using a real-time polymerase chain reaction (PCR). We found that the null GSTM1 and GSTT1 genotypes were associated with an increase in the risk of developing coronary heart disease (OR = 1.14; 95% CI: 0.71 - 1.82; OR = 1.38; 95% CI: 0.82 - 2.32), respectively, but this increase was not significant. Patients who smoke having the null genotypes of GSTM1 (OR: 1.63 (1.10 - 2.63)) and GSTT1 (2.66 (1.50 - 4.72)) and both (3.20 (1.37 - 7.45)) were at a higher risk for developing coronary heart disease. In conclusion, the finding of a significant association between GSTM1 and T1 with smoking status may influence cardiovascular disease via DNA damage.

Age-related macular degeneration is the leading cause of legal blindness in the developed world, and yet its pathogenesis remains poorly understood. Oxidative stress may play a major role in the etiology and pathogenesis of age-related disorders such as age-related macular degeneration. Catalase is an antioxidant enzyme which plays an important role in the detoxification of free oxygen radicals. Malondialdehyde is a marker that shows free radical damage. We have measured the erythrocyte activity of catalase and the serum level of malondialdehyde in 30 patients with age-related macular degeneration and 60 healthy subjects. Patients with age-related macular degeneration showed significantly lower catalase activity compared to healthy subjects (p = 0.002). Plasma malondialdehyde level of the patient group was significantly higher than that of the controls (p = 0.038)., (Copyright 2004 S. Karger AG, Basel)

Glutathione S-transferase (GST) enzymes play an important role in drug metabolism. GST is a multigene family of enzymes involved in the detoxification and in a few instances activation of a wide variety of chemicals. Detoxification features make it plausible to search for GST polymorphism in patients with drug eruption. The GSTM (mu), GSTT (theta) and GSTP (pi) have been shown to be polymorphically distributed. The GSTT1, GSTM1 and GSTP1 gene polymorphism were detected using real-time PCR. GSTM1 and GSTT1 null genotypes were found to be associated with an increased risk of drug eruption (OR 2.27, 95% CI 1.20-5.21; OR 2.48, 95% CI 1.12-6.39, respectively). No relationship was observed between the null combination of the GSTM1 and GSTT1 genotype polymorphisms and drug eruption risk (OR 2.65, 95% CI 0.62-11.25). Our results show that GSTP1 polymorphism is not a significant contributor to drug eruption risk. The GSTM1 and GSTT1 gene polymorphisms seem to be associated with the development of drug eruption. Further studies may shed additional light on the role of GSTM1, GSTT1 and GSTP1 in drug eruption.

Since oleic acid (OA) induces morphologic and cellular changes similar to those observed in human acute lung injury (ALI) and acute respiratory distress syndrome, it has become a widely used model to investigate the effects of several agents on pathogenesis of lung injury. The antioxidant and anti-inflammatory properties of N-acetylcysteine (NAC) has been documented in many lung injury models. In this study, we evaluated the role of NAC in an OA-induced lung injury model by measuring myeloperoxidase (MPO) activity, malondialdehyde (MDA) and 3-nitrotyrosine (3-NT) levels in lung tissue. Five groups labelled Sham, NAC, OA, Pre-OA-NAC and Post-OA-NAC were determined. ALI was induced by intravenous administration of OA. The pre-OA-NAC group received iv NAC 15 min before OA infusion and the post-OA-NAC group received iv NAC 2 h after OA infusion. In both of the NAC treatment groups' blood and tissue samples were collected 4 h after OA infusion, independent from the time of NAC infusion. The MPO activity, MDA and 3-NT levels in lung homogenates were found to be increased in OA group and the administration of NAC significantly reduced tissue MPO, MDA and 3-NT levels (p = 0.0001) Lung histopathology was also affected by NAC in this OA-induced experimental lung injury model.